iFyber provides a suite of biofilm tests to verify the efficacy of antimicrobial medical devices and drugs. Biofilms are complex communities of microbes protected within a matrix of sugars, proteins, and extracellular DNA. These characteristics, in addition to their tolerance to antibiotics, make biofilms significantly more difficult to prevent and remove. Research has shown that between 60 to 90% of chronic wounds are associated with biofilms. The development of technologies to combat resistant biofilm infections often requires custom biofilm test models. Biofilm tests at iFyber can be tailored to evaluate technologies in a way that meets the client’s objective.

We focus our biofilm testing on determining the efficacy of new technologies and actively developing novel assay systems based on our client’s specific needs. Initial in vitro test methods can be used to screen formulations and measure antimicrobial and anti-biofilm efficacy for a range of applications, from therapeutics to antifouling technologies. For wound care applications, our ex vivo model allows for the evaluation of prototype formulations on a more clinically relevant matrix before the start of costly in vivo animal studies – saving money and reducing time to market. In addition to our standard and custom biofilm testing program, iFyber also offers a host of antimicrobial susceptibility testing options for planktonic (free-floating) bacteria. Biofilms continuously shed planktonic bacteria that can disperse and attach to other areas of the wound, potentially forming new biofilms. By taking a multi-method biofilm testing approach, treatments can be evaluated for their ability to both prevent the formation of biofilms and eradicate existing biofilms.

Examples of relevant materials for biofilm testing include new antibiotics/biocides, coatings, nanomaterials, wound dressings, coated natural tissue matrices, and coated medical devices.

 

Biofilm Testing

  • MBEC in vitrobiofilm test system
  • Microwell biofilm susceptibility and eradication assays
  • Porcine dermal explant ex vivomodel
  • Artificial sputum in vitromodel of cystic fibrosis
  • Modified Robbins device

Antimicrobial Susceptibility Testing

  • Dynamic contact assay
  • Time-kill assay
  • Zone of inhibition assay
  • Time course of antimicrobial activity
  • MIC assay
  • MTT assay
  • Bacterial selective plating
  • Bacterial attachment

Histology & Imaging

  • Simple staining for bacteria (e.g., methylene blue, safranin, crystal violet)
  • Differential staining (e.g., Gram staining, GMS staining)
  • LIVE/DEAD staining
  • Confocal microscopy and 3D reconstruction for ex vivo model
    • Visualization of bacteria using GFP or luciferase (lux) reporter systems

Contact us today to discuss your application-specific antimicrobial and biofilm testing needs.

 

Biofilm Testing Labs

Our BSL-2 biofilm testing facilities are fully equipped to handle the contract testing needs of medical device and pharmaceutical companies.  Located in Ithaca, New York, our lab is GLP compliant and has a menu of clinically relevant bacterial species to grow and evaluate biofilm test models.

What is a Biofilm?

A biofilm consists of bacterial communities encased within a self-produced protective matrix of sugars and proteins. Compared to free-floating (planktonic) forms of bacteria, biofilm-associated bacteria are much more difficult to kill due in large part to the presence of this protective matrix and a reduction of metabolic activity. Biofilms have been demonstrated to form on a range of surfaces, including medical devices and human tissue systems. iFyber offers a menu of biofilm tests and assays that assess biofilm prevention and dispersal efficacy.

Why Include Biofilm Susceptibility Testing?

Biofilms pose a major clinical threat. Current estimates from the US National Institutes of Health suggest biofilms account for over 80% of all microbial infections. Given the relatively recent understanding of biofilms, biofilm testing is becoming an increasingly common addition to the screening profile of new antimicrobial technologies.

Ex Vivo Porcine Dermal Model for Mature Biofilms

What is it?

This custom biofilm test method is a robust model of a mature bacterial biofilm, which is a hallmark of many bacterial soft tissue infections. This model isolates the biofilm component of these infections through the growth of a mature biofilm within a natural porcine dermal matrix. The resulting biofilm (typically grown over 72 hours) exhibits high tolerance to many antibiotics and, importantly, incorporates a relevant substrate when assessing product efficacy in the presence of soft tissue. iFyber has extensive biofilm testing experience adapting this model to fit our client's needs. Specifically, this biofilm test can be used to evaluate antimicrobial and anti-biofilm activity of a technology against a model of mature biofilm that exhibits tolerance to high doses of antibiotics. In this model, tissue explants are created, wounded, and subsequently infected to enable high-throughput screening using a number of assay formats (e.g., prevention, eradication, dispersal, synergy).

 

Why use it?

The characteristics of biofilms grown on non-biological substrates can differ from what occurs in soft tissue. When using our biofilm test methods for evaluating antimicrobial solutions to eradicate biofilms from wounds, it can be informative to use a biofilm assay that closely approximates the wound environment. Porcine skin is physiologically very similar to that of human skin. Using skin as both the substrate for attachment and the primary source of nutrition allows for the development of a biofilm that mimics the characteristics of biofilms found in human wounds. This ex vivo model biofilm test allows for high throughput screening of compounds, prototype formulations, solid wound dressings, etc., to complement/augment more costly animal studies.

Minimum Biofilm Eradication (MBEC) Assay

What is it?

This high throughput assay comes in a microwell plate format, enabling growth of 96 bacterial biofilms on polystyrene pegs that protrude down from the lid of the plate into the corresponding well. As the biofilms are attached to the removable peg lid, this assay is set up to allow rapid biofilm testing of a range of materials for anti-biofilm efficacy. Quantification is achieved by recovering remaining bacteria from the peg lid and enumerating either by standard agar plate counts, or for high throughput purposes, by optical density. iFyber has extensive biofilm testing experience using this assay system to screen anti-biofilm technologies against both single-and multi-species biofilms.

 

Why use it?

The MBEC assay system (also known as the Calgary Biofilm Device) is accepted as a standard anti-biofilm testing approach by the American Society for Testing and Measurement (ASTM) and is widely used in both industrial and academic settings. In this model, biofilms are cultured on a removable peg lid to provide a high throughput screening tool for assessing biofilm dispersal, eradication, or prevention. The system can be used to test both soluble and insoluble materials, unlike other microwell plate-based assay systems.

Microwell Biofilm Susceptibility Assays

What is it?

This 96-well microtitre plate-based assay involves culturing bacterial biofilms in the wells of the plate, then exposing the formed biofilms to the test agent for a pre-defined time. The total remaining biofilm after exposure to the antimicrobial material is typically quantified by fluorescence methods using the nucleic acid stain, SYTO-9, or colorimetrically using Crystal Violet — a visible Gram stain that labels all components of a biofilm.

Why use it?

This assay is often used as an initial screen for anti-biofilm technologies. It is quick and easy to run and can help guide further biofilm testing using more complex model systems. The fluorescent dye strategy allows for the quantification of biofilm bacteria or total biomass, which includes the exopolysaccharide matrix.